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Modifications in Adenoviral Coat Fiber Proteins and Transcriptional Regulatory Sequences Enhance Transgene Expression
HARRIS PERLMAN, HONGTAO LIU, CONSTANTINOS GEORGANAS, JAMES M. WOODS, M. ASIF AMIN, ALISA E. KOCH, THOMAS WICKHAM, IMRE KOVESDI, TOSHIAKI MANO, KENNETH WALSH, and RICHARD M. POPE
ABSTRACT.
Methods. Primary human fibroblasts and macrophages were infected with standard replication-defective adenoviruses, adenoviral vectors containing modified fiber coat proteins expressing Arg-Gly-Asp (RGD) or heparin sulfate binding moieties, or a tetracycline-regulatable transgene transcription system. Each of these vectors expressed the ß-galactosidase gene (ß-Gal), which was quantified by flow cytometry. Ankle joints from rats with adjuvant induced arthritis were transduced intraarticularly with each of the vectors and ß-Gal expression was quantified by flow cytometry. Results. Primary human fibroblasts and macrophages displayed marked increases in transgene expression from both modified fiber protein vectors and from the tetracycline-regulatable vector, compared to an unmodified vector expressing the transgene from the cytomegalovirus promoter/enhancer. In the rat model, the modified fiber protein vectors and the tetracycline-regulatable vector system also displayed increased transgene expression in inflamed rat joints. Conclusion. Adenovirus attachment and uptake by cells and promoter strength limit transgene expression from conventional adenoviral vectors in models of rheumatoid arthritis. (J Rheumatol 2002;29:1593-600) Key Indexing Terms:
RHEUMATOID ARTHRITIS
From the Division of Rheumatology, Northwestern University Medical School; VA Chicago Heathcare System, Lakeside Division, Chicago, Illinois; GenVec, Inc., Gaithersburg, Maryland; Division of Cardiovascular Research, Molecular Cardiology, Boston University School of Medicine, Boston, Massachusetts, USA. Supported by a Northwestern Memorial Foundation grant and National Institutes of Health grant AR02147 to Dr. Perlman; National Institutes of Health grants AR43642, AR30692, and AR62229 and a grant from the Arthritis Foundation to Dr. Pope; and by National Institutes of Health grants AR41492 and HL58695, funds from the VA Research Service, and Gallagher Professorship for Arthritis Research to Dr. Koch. H. Perlman, PhD; H. Liu, MD, PhD; C. Georganas, MD (current address: Rheumatology Department, 251 Hellenic Airforce V.A. General Hospital, Athens, Greece); J.M. Woods, PhD; M.A. Amin, MD; A.E. Koch, MD; R.M. Pope, MD, Division of Rheumatology, Northwestern University Medical School, and VA Chicago Heathcare System; T. Wickham, PhD; I. Kovesdi, PhD, GenVec, Inc.; T. Mano, MD; K. Walsh, PhD, Division of Cardiovascular Research, Molecular Cardiology, Boston University School of Medicine. Address reprint requests to Dr. R.M. Pope, Division of Rheumatology, Northwestern University Medical School, Ward 3-315, 303 East Chicago Ave., Chicago, IL 60611. E-mail: RMP158@northwestern.edu Submitted August 13, 2001; revision accepted January 15, 2002. |